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LAMP (loop-mediated isothermal amplification) primer and detection method for detecting porcine circovirus 3

A technology of porcine circovirus and detection method, applied in the field of LAMP detection of porcine circovirus type 3

Inactive Publication Date: 2017-12-19
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve the problem of the loop-mediated isothermal amplification detection method for porcine circovirus type 3 in the prior art, the application discloses a LAMP primer for detecting porcine circovirus type 3

Method used

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  • LAMP (loop-mediated isothermal amplification) primer and detection method for detecting porcine circovirus 3
  • LAMP (loop-mediated isothermal amplification) primer and detection method for detecting porcine circovirus 3
  • LAMP (loop-mediated isothermal amplification) primer and detection method for detecting porcine circovirus 3

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Embodiment 1

[0028] 1 Materials and methods

[0029] 1.1 Viruses, recombinant plasmids and disease materials

[0030] Viruses: PCV3, PCV1, PCV2a, PCV2b, porcine pseudorabies virus (PRV), swine fever virus (CSFV), porcine epidemic diarrhea virus (PEDV), etc. are all isolated and preserved by our laboratory.

[0031] Recombinant plasmid: pMD19-T-Cap, containing PCV3ORF2 gene, the concentration is 1.89×10 -7 ng / μL, the gene copy number is 5.22×10 10 copies / μL, constructed and provided by Nanjing Gensirui Biotechnology Co., Ltd.

[0032] Pig clinical samples: from 38 pig farms in Jiangsu, Shandong, Zhejiang and Fujian provinces from 2016 to 2017, clinically showing respiratory symptoms, 68 samples of lymph node and lung tissue. Store at -20°C for later use.

[0033] 1.2 Viral nucleic acid extraction

[0034] Extract according to the instructions of the DNA extraction kit, use the method of the TRIZOL reagent instructions to extract the viral nucleic acid, and store at -20°C.

[0035] 1.3...

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Abstract

The invention relates to the technical field of virus detection and especially relates to an LAMP (loop-mediated isothermal amplification) primer for detecting porcine circovirus 3. The LAMP primer is shown by a sequence 1-6 in a sequence list. The LAMP primer is utilized to perform amplification; if an amplification result is obtained, a sample to be detected contains the porcine circovirus 3; if no amplification result is obtained, the sample to be detected has no the porcine circovirus 3. The method has the advantages of sensitivity, specificity, convenience, quickness and application to PCV3 detection and clinical diagnosis.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a LAMP primer for detecting porcine circovirus type 3, and also relates to a LAMP detection method for porcine circovirus type 3. Background technique [0002] Porcine circovirus (PCV) is a member of the genus Circovirus in the family Circoviridae. The virus has two serotypes, PCV1 and PCV2. PCV2 infection can cause a variety of clinical diseases such as multisystemic wasting syndrome (PMWS), porcine dermatitis nephropathy syndrome (PDNS), porcine respiratory tract syndrome (PRDC), sow reproductive failure syndrome, diarrhea and congenital tremor in weaned piglets. It has brought huge economic losses to the global aquaculture industry. However, the etiology of porcine dermatitis nephrotic syndrome (PDNS) has been controversial. In recent years, American scholars first discovered that there is a new porcine circovirus in the tissues of reproductive disorder sows, PDNS, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2563/107
Inventor 白娟姜辰龙姜平张日腾李勇严秀文
Owner NANJING AGRICULTURAL UNIVERSITY
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